期刊
JOURNAL OF FUNGI
卷 7, 期 4, 页码 -出版社
MDPI
DOI: 10.3390/jof7040319
关键词
Candida; aequorin; calcium homeostasis; alkaline stress; amiodarone
资金
- Swiss National Research Foundation [31003A_172958]
- Swiss National Science Foundation (SNF) [31003A_172958] Funding Source: Swiss National Science Foundation (SNF)
In Candida albicans, calcium ions play a crucial role in regulating signaling pathways, cell polarization, and hyphal extension. Aequorin-based Ca2+ sensing system provides a valuable tool for studying Ca2+ homeostasis in this fungal pathogen. Mutants in key Ca2+ channels showed reduced response to alkaline stress, highlighting the importance of these channels in Ca2+ uptake and homeostasis.
In Candida albicans, calcium ions (Ca2+) regulate the activity of several signaling pathways, especially the calcineurin signaling pathway. Ca2+ homeostasis is also important for cell polarization, hyphal extension, and plays a role in contact sensing. It is therefore important to obtain accurate tools with which Ca2+ homeostasis can be addressed in this fungal pathogen. Aequorin from Aequorea victoria has been used in eukaryotic cells for detecting intracellular Ca2+. A codon-adapted aequorin Ca2+-sensing expression system was therefore designed for probing cytosolic Ca2+ flux in C. albicans. The availability of a novel water-soluble formulation of coelenterazine, which is required as a co-factor, made it possible to measure bioluminescence as a readout of intracellular Ca2+ levels in C. albicans. Alkaline stress resulted in an immediate influx of Ca2+ from the extracellular medium. This increase was exacerbated in a mutant lacking the vacuolar Ca2+ transporter VCX1, thus confirming its role in Ca2+ homeostasis. Using mutants in components of a principal Ca2+ channel (MID1, CCH1), the alkaline-dependent Ca2+ spike was greatly reduced, thus highlighting the crucial role of this channel complex in Ca2+ uptake and homeostasis. Exposure to the antiarrhythmic drug amiodarone, known to perturb Ca2+ trafficking, resulted in increased cytoplasmic Ca2+ within seconds that was abrogated by the chelation of Ca2+ in the external medium. Ca2+ import was also dependent on the Cch1/Mid1 Ca2+ channel in amiodarone-exposed cells. In conclusion, the aequorin Ca2+ sensing reporter developed here is an adequate tool with which Ca2+ homeostasis can be investigated in C. albicans.
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